乳双歧杆菌Probio-M8连续传代过程中遗传稳定性分析  

GENETIC STABILITY ANALYSIS OF BIFIDOBACTERIUM ANIMALIS SUBSP.LACTIS PROBIO-M8 DURING CONTINUOUS PASSAGE CULTURE

作　　者：刘晓晔 刘凯龙 李伟程 刘佳琪 张和平[1] LIU Xiao-ye;LIU Kai-long;LI Wei-cheng;LIU Jia-qi;ZHANG He-ping(Key Laboratory of Dairy Biotechnology and Engineering,Ministry of Education,Key Laboratory of Dairy Products Processing,Ministry of Agriculture and Rural Affairs,Inner Mongolia Key Laboratory of Dairy Biotechnology and Engineering,1nner Mongolia Agricultural University,Hohhot 010018,China)

机构地区：[1]内蒙古农业大学乳品生物技术与工程教育部重点实验室,农村部奶制品加工重点实验室,内蒙古乳品生物技术与工程重点实验室,呼和浩特010018

出　　处：《营养学报》2023年第5期504-511,520,共9页Acta Nutrimenta Sinica

基　　金：国家现代农业产业技术体系,内蒙古科技重大专项(No.2021ZD0014)。

摘　　要：目的探究乳双歧杆菌Probio-M8(Bifidobacterium animalis subsp.lactis Probio-M8)在连续传代中的遗传稳定性。方法厌氧环境下,乳双歧杆菌Probio-M8在胰胨植胨酵母膏琼脂培养基(trypticase-phytone-yeast extract agar medium,TPY培养基)中连续传100代时,对不同代时菌株(0、25、50、75、100代)的菌落形态变化、碳水化合物利用情况、细菌代谢物产生情况及比较基因组学进行分析。以乳双歧杆菌Probio-M8原始菌株基因组作为参考序列,利用二代数据对三代数据识别到的突变位点进行验证。结果与原始菌株相比,不同代时菌株基因组大小及GC含量均无显著差异(P>0.05)。两两菌株间ANI值均大于99%。从不同代时15株菌的基因组中共鉴定到31个SNP位点,包括7个同义突变和7个非同义突变,其余17个位于基因间区。每株菌的SNP个数均小于21,且每两个代时菌株间突变位点不会稳定存在。同时,在连续传代100代时过程中不同代时菌株的菌落形态、碳水化合物利用情况和细菌代谢物产生情况等均无显著变化。结论乳双歧杆菌Probio-M8在厌氧环境的TPY培养基中遗传稳定性良好,为该菌株开发应用提供理论参考。Objective To investigate the genetic stability of Bifidobacterium animalis subsp.lactis Probio-M8(BK animalis subsp.lactis Probio-M8)during dontinuous passage culture.Methods Under anaerobic environment,when Probio-M8 was continuously cultured for 100 passage in trypticase-phytone-yeast(TPY)extract agar medium,the morphological changes of bacterial colonies,carbohydrate utilization,bacterial metabolite production and comparative genomics in different generations(0、25、50、75、100 generations)were analyzed.The original genome of BK animalis subsp.lactis Probio-M8 was used as a reference sequence.The second-generation data were used to verify the mutation sites identified by the third-generation data.Results Compared with original strain,there were no significant differences in genome size and GC content among different generations(P>0.05).The ANI values between each two strains were all greater than 99%.A total of 31 SNPs were identified from 15 strains,including 7 synonymous mutations and 7 non-synonymous mutations,and the other 17 SNPs were located in the intergene region.The number of SNP in each of the other strains was less than 21,and they could not exist stably during passage culture.Meanwhile,there were no significant differences in colony morphology,carbohydrate utilization and bacterial metabolites of different generations in the passage process.Conclusion The genetic stability of BK animalis subsp.lactis Probio-M8 in the TPY medium in anaerobic environment is relatively good,which provides a theoretical reference for the development and application of this strain.

关 键 词：乳双歧杆菌Probio-M8 连续传代 胰胨植胨酵母膏琼脂培养基 遗传稳定性 

分 类 号：TS201.3[轻工技术与工程—食品科学]